Identification to the species level of <Emphasis Type="Italic">Lactobacillus</Emphasis> isolated in probiotic prospecting studies of human,animal or food origin by 16S-23S rRNA restriction profiling

Background  

The accurate identification of Lactobacillus and other co-isolated bacteria during microbial ecological studies of ecosystems such as the human or animal intestinal tracts and food products is a hard task by phenotypic methods requiring additional tests such as protein and/or lipids profiling.     

Morphological differences among egg nests and adult individuals of Cicadatra persica (Hemiptera,Cicadidae), distributed in Erneh,Syria

The aim of this study is determining the different patterns of egg nests and the morphological differences between the specimens of Cicadatra persica Kirkalidy, 1909 (Hemiptera: Cicadidae) distributed in fruit orchards in Erneh located on AL-Sheikh mountain south west of Syria. The appearance of 80 egg nests was studied, and the results showed that there were two basic patterns of egg nests laid by Cicadatra persica, 90% of the egg nests were of the first pattern (consists of several adjacent slits), while 10% of them were of the second pattern (consists of several divergent slits). A random sample consisting of 300 specimens (150 males and 150 females) were also studied concentrating on the differences in the color of the supra-antennal plate and in the number of spurs on the tibia of the hind legs. The results showed that there were two basic patterns of individuals based on the differences in the color of supra-antennal plate. The first pattern (individuals with yellow supra-antennal plates), constituted more than 90%, and the second one (individuals with black supra-antennal plates) constituted less than 10%. The results also showed that there were 27 different patterns based on the number of spurs on the tibia of the hind legs. One of them was a common pattern (2, 3) whose individuals have 2 spurs on the upper side of the tibia of the hind legs and 3 spurs on the lateral side of the tibia of the hind legs. The total percent of this common pattern was 76%. The other 26 patterns were different from each other, and the total percent of all these different patterns was 24%.     

Collagenase predigestion on paraffin sections enhances collagen immunohistochemical detection without distorting tissue morphology

Reliable immunohistochemical detection of collagen in formalin fixed, paraffin embedded tissues requires protease digestion. While these pan-proteases (pepsin, trypsin, protease K, etc.) enhance collagen detection, they also digest many other tissue proteins and produce poor cellular morphology and unrecognizable cellular structures. Balancing the conditions (protease type, concentration, incubation time and temperature) to digest some, but not all, proteins in a tissue section while optimizing collagen detection requires one to compromise improved collagen immunolabeling with adequate cellular morphology. Furthermore, optimal conditions for digesting tissue proteins to enhance collagen detection vary among tissue types and their fixation. Although brain is not typically subject to these deleterious consequences, structures such as epithelium, spermatids, stroma etc. and other tissues with complicated histology are profoundly affected. To resolve this technical dilemma, we discovered a novel use for collagenase to enhance collagen immunodetection without affecting the noncollagen proteins, thereby preserving tissue morphology. Collagenase, which is typically used in vitro for disassociation of cells, has never been used reliably on formalin fixed, paraffin embedded tissue sections. This new use of collagenase for immunohistochemistry promotes increased collagen immunolabeling, is easy to use, is versatile, and allows preservation of tissue structure that provides maximal and accurate histological information.     

Molecular Dynamics Simulation Study of the Interaction of Piscidin 1 with DPPC Bilayers: Structure-Activity Relationship

Abstract

To study structure-activity relationship of antimicrobial peptides and to design novel antimicrobial peptides with selectivity for bacterial cells, we have performed molecular dynamics simulations of the interaction of Piscidin (Pis1) and its two analogues (Pis1-AA and Pis1-PG) with dipalmitoylphosphatidylcholine (DPPC) bilayer through 45 ns. Our results inform us of the detailed location and orientation of the peptide with respect to the bilayer as well as provide about hydrogen-bond-formation patterns and electrostatics interactions. Simulations show that Pis1 and Pis-AA form the most hydrogen bonds and Pis-PG forms the fewest hydrogen bonds with lipid. Thus, Pis1 and Pis-AA should have stronger interactions with the lipid head group when compared to Pis-PG. Experimental studies have shown that Pis1 and Pis1-AA have a high antimicrobial and hemolytic activities, and Pis1-PG has low hemolytic activity while keeps potent antimicrobial activity. Our results complement the previous experimental studies. According to our MD results and previous experimental studies, Pis1 and Pis1-AA are more effective at the zwitterionic bilayer comparing Pis1-PG. These properties of Pis1-PG could be accordance with its low hemolytic activities.     

Life history parameters of the coccinellid beetle, Oenopia conglobata contaminata, an important predator of the common pistachio psylla, Agonoscena pistaciae (Hemiptera: Psylloidea)

The predatory beetle, Oenopia conglobata contaminata, is associated with the common pistachio psylla, Agonoscena pistaciae, the major pistachio pest in Iran. Successful development and reproduction on both A. pistaciae and Aphis gossypii (the major weed aphid in the pistachio orchards) indicated they were suitable prey for O. conglobata contaminata. Under ample prey supply, larval development on A. pistaciae was shorter and mortality was lower compared to those reared on A. gossypii. Furthermore, this ladybird attacks and destroys a large number of psyllid nymphs during the whole of its larval period (620±17 fourth stage nymphs) and also in its adult stage (191±7.5 4^th stage nymphs daily). The optimum temperature for development was 30°C, the theoretical threshold for development was 13°C and thermal requirements from egg to adult was estimated as 196 degree-days (°D). Fecundity for the first 21 days of adult life was 387 and 355 eggs when females fed on A. pistaciae and A. gossypii nymphs, respectively. The intrinsic rate of increase (r_m) at 27.5°C was 0.19 and 0.18 when ladybirds were fed on psyllid and aphid nymphs, respectively. In a laboratory choice experiment, the adult ladybirds showed a strong preference for A. pistaciae compared to A. gossypii. This was in agreement with our field observation that O. conglobata contaminata is almost always found on psyllid colonies, even when aphids were plentiful on weeds in the pistachio orchards. The influence of the predator on psyllid seasonal population under natural condition was not studied in this investigation and this subject must be measured in subsequent trials.     

Investigation of apoptosis in cultured cells infected with equine herpesvirus 1

Many viruses alter different stages of apoptosis of infected cells as a strategy for successful infection. Few studies have addressed mechanisms of equine herpesvirus 1 (EHV-1) strain-induced cell death. We investigated the effect of an abortigenic strain (AR8 strain) on heterologous Madin–Darby bovine kidney cells and homologous equine dermis (ED) cells cell lines. We compared morphologic and biochemical features of early and late apoptosis at different postinfection times. We investigated translocation of phosphatidylserine to the cell surface, nuclear fragmentation and changes in the cytoskeleton using flow cytometry and annexin V/propidium iodide staining, DNA laddering, terminal deoxynucleotidyl transferase UTP nick-end labeling assay and immunofluorescence staining of cytokeratin 18 cleavage. AR8 EVH-1 strain interfered with apoptosis in both cell lines, particularly during the middle stage of the replication cycle; this was more evident in ED cells. Although this antiapoptotic effect has been reported for other alpha herpesviruses, our findings may help elucidate how EHV-1 improves its infectivity during its cycle.     

The Influence of Host Species on Some Biological and Behavioural Aspects of Dibrachys boarmiae (Hymenoptera: Pteromalidae), Parasitoid of Kermania pistaciella (Lepidoptera: Tineidae)

Dibrachys boarmiae was recently found as a parasitoid of the pistachio twig borer moth, Kermania pistaciella, an important pistachio pest in Iran. Several biological and behavioural aspects of the wasp were studied under controlled conditions using three different lepidopterous host species, K. pistaciella, Sitotroga cerealella and Galleria mellonella. Development time from egg to adult decreases from 54 days at 15°C to 12.6 days at 32.5°C. The thermal constant was estimated 270°D, and the lower development threshold was at 9.9°C. Longevity of females that had the opportunity to oviposit and access to food (honey) was on average 23 days, and 134 offspring were produced during 13 days of oviposition. Ninety percent of ovipositions occur during the first 10 days of the parasitoid's life, although no pre-oviposition period was found. If females were deprived of food, longevity as well as fecundity, were drastically reduced to 8 days and only 79 offspring respectively. Females were able to mate immediately after emergence and the sex ratio of the progeny was strongly female biased on all examined hosts, especially on the largest host G. mellonella (0.08 male). When females were kept with a male throughout their life, the total number of progeny decreased to 92 wasps on average but the sex ratio was unaffected. The parasitoid significantly preferred to attack the pre-pupal stage, and this influenced the size of clutches allocated to different host stages. The parasitoid laid larger clutches on bigger hosts.     

Stent implantation through a self-expanding stent

Jailing of a side-branch is a known complication of stent implantation, and makes access to the side-branch difficult, especially if the stent is of the self-expanding type. Although plain balloon angioplasty is feasible for the jailed side-branches, the use of newer devices (a stent, Rotablation or atherectomy) has not been described. We describe a novel way of treating a side-branch jailed by a self-expanding stent by using stent implantation through the strut of a self-expanding stent.